Abstract
摘要:在癌症治療的重要挑戰之一,是即使是患有相似腫瘤的病人,對相同的化療藥物卻仍有不同的治療反應。另外,不同的化療藥物治療效果或毒性影響往往因人而異。因此發展一套能滿足病人客製化,個人化需求的體外化療藥物篩選方法及檢測方式是重要且有價值的。
本計畫為期三年,主要目標為利用微流檢測方式,提供乳癌病人及血癌病人個人化,客製化的化療需求。本計畫將建立三種檢測工具:1.細胞微陣列之微流平台;2.碟片形式之微流檢測及3.單細胞之微流檢測平台。每年的計畫目標為︰
第一年: 透過測試具有代表性的細胞株以驗証各種微流檢測構想。這階段涉及建立處理癌細胞株的設備,以供細胞實驗使用,並藉細胞株驗證和最佳化三種微流檢定法。將用細胞微陣列平台進行乳癌細胞株對於化療藥物敏感性之測試。為了評估化療效果的好壞,擬建立一碟片形式的檢測平台,利用該平台能有效的計數出血液中循環腫瘤細胞數目,做為評估治療效果的參考指標。在本年度,我們先用細胞株進行模擬測試。對血癌細胞株,以單細胞陣列排列細胞(大約10至100個細胞)來測試單一細胞的化療反應。也會開始比較不同平台的測試結果。
第二年: 直接從乳癌病人身上取出初代細胞(primary cell)進行實驗,並建立該試驗規則,並建立處理初代細胞的設備以支援相關的實驗。在本年度,將從乳癌腫瘤切片中獲得初步細胞,而後利用細胞微陣列平台進行化療藥物敏感性測試。另外,將用碟片平台,檢測病人化療前、後血液中循環腫瘤細胞(CTC)的變化。這個結果會和一既存方法CellSearch® (Veridex Inc., FDA核可)比較。這些在化療過程難以清除的細胞可能具有類似幹細胞的性質。本計畫將利用前述的抓取單一細胞之微陣列平台(若細胞量較多,則採用細胞微陣列平台),研究這些單一細胞的化療敏感特性及分生特性。
第三年: 為延續前兩年的努力結果,將對乳癌病人實施上述的微流檢測,以建立個人化、客製化的的化療藥物敏感性測試及化療效果的評估,並將實際支援台大醫院的臨床需求。對各個檢定法將選適量病人數進行測試。這些微流檢測的結果具有潛力做為乳癌化療後的預後評估指標。
本團隊成員包括台大乳房外科的黃俊升醫生,乳房腫瘤科的林璟宏醫生,專精於醫檢分生診斷之林亮音教授,自羅氏藥廠退休,專精於個人化醫療的外藉專家楊鎣博士,專精於細胞培養的細胞生物學家李心予副教授及微流專家胡文聰教授。我們相信團隊的專業能力應可確保計畫的成功。
預期成果:(1) 建立一最佳化的微流檢測方式,做為輔助治療乳癌之臨床工具。(2)在病人化療前提供病人客製化的體外化療藥物測試,並追蹤治療後的效果,以達到最好的預後結果。(3) 了解收集到的稀少細胞(例如,血液中循環的腫瘤細胞)的分生特性。
Abstract: The selection of chemotherapeutic drugs used in cancer therapy is currently based on clinical trial results to show superior overall response of one drug over others. However, tumors that are clinically similar vary in their response to the same drug. Because of wide variations of drug efficacy and significant toxicity of most chemotherapeutic drugs, it would be of obvious value to develop an in vitro assay that can identify optimal therapies for the individual patients, or personalized medicine.
The overarching goal of this three-year project aims to provide patient-specific approach to address specific chemotherapeutic needs in breast cancer patients using microfluidics assays. Three such assays will be established: cellular microarray microfluidic assay, disk-based microfluidic assay and single cell microfluidic assay. Yearly goals are as follows:
Year 1: Proof of concept of various microfluidic assays via testing over a range representative cell lines. This involves establishing a cancer cell line core facility that provides support for various cellular testing, as well as overall validation and optimization of the three microfluidic assays for chemotherapeutic drug selection and efficacy via the cell lines. Chemosensitivity studies for breast cancer cell lines will be tested in the cellular microarray assay. Chemotherapeutic efficacy testing will be established in the disk-based assay based on enumeration of breast circulating tumor cells (CTC) in blood – simulated by cell lines in Year 1.
Year 2: Establish testing protocol via primary cells from breast cancer. This involves establishing a primary cell core facility to support a wide range of tests. Cells from breast tumor biopsy will be tested for chemosensitivity in the cellular microarray assay. Blood sample pre- and post-chemotherapy will be tested for circulating tumor cell (CTC) via the disk-based assay. Results will be compared to that by CellSearch® (Veridex Inc., FDA approved).
Year 3: Perform patient-specific drug chemosensitivity and chemotherapeutic efficacy testing via microfluidic assays for breast cancer to partially support clinical needs in the National Taiwan University Hospital. This entails leveraging on efforts made on the previous two years. Suitable number of patients will be chosen for each study. Disease prognosis will also be made as evidence of the potential contribution of the microfluidics assays in the chemotherapy of the two types of cancer.
The PIs in this project is believed to be highly qualified for the tasks at hand. The team consists of a breast surgeon (CS Huang), a breast oncologist (CH Lin), a molecular diagnosis specialist in laboratory medicine (LI Lin), an international expert in personalized medicine retired from Roche Pharmaceutical (P Lau), a cell biologist with specialty in cell culture (HY Lee) and a microfluidics specialist (AM Wo). Together, we believe our expertise should ensure success of this project.
Expected results are as follows: (1) establish and fine-tune the optimal operational characteristics of the microfluidics assays as clinical tools in the fight for breast cancer; (2) provide patient-specific in-vitro chemotherapeutic drug testing prior to therapy and track its post-treatment efficacy to maximize disease prognosis; (3) molecular characterization of collected rare cells, e.g. CTC cells.
Keyword(s)
微流體
癌症
化療
乳癌
循環腫瘤細胞
幹細胞
醫療儀器
Microfluidics
chemotherapy
drug selection
breast cancer
circulating tumor cell
stem cell
commercialization
medical device.